Hi,
we´ve been trying to determine whether a bacterial intron is splicing
also in eucaryotes. Therefore we first modified vector pEGFP-N1 by
deleting the ATG codon and the parts of the Kozak sequence of the EGFP
gene. We then planned to introduce a new startcodon and a Kozak sequence
5´ to the intron and look for expression of the EGFP after transfection.
However when we tested the modified pEGFP-N1 vector (the one without ATG
and Kozak seq), we already got fluoresence. Thus apparently the EGFP is
produced in eucaryotic cells eventhough the ATG is missing.
Does anyone have an explanation for this? Or has anyone had the problem
as well??
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